The effect of blood collection methods on the expression of monocyte cellular adhesion molecules by Erin L. Skornicka.

Abstract

Cellular adhesion molecules (CAMs) play an important role in the inflammatory response by allowing white blood cells to adhere to the endothelium and then migrate into infected tissues. The goal of this study is determine the effect of blood collection techniques on the ability to detect changes in these molecules. In this study, the effects of time (0 hour versus 2 hours), temperature (room temperature versus 4°C), and anticoagulants (EDTA, potassium oxalate, sodium citrate, and heparin) were examined to see which blood collection techniques retained accurate results when compared to samples that were processed immediately. Cells, both unstimulated and stimulated with PMA, from healthy volunteers were labeled with various antibodies against CAMs (anti-ICAM-l, LFAlbeta, L-selectin, Mac-l, LFA-lalpha, and VLA-4) followed by FITC-Iabeled goat anti-mouse antibody then analyzed using a flow cytometer. From these experiments, the time of incubation and temperature of storage seem to influence the expression of CAMs. The anticoagulant does not seem to play an important role in altering CAM expression. In these studies, the best storage medium for monocytes when examing Mac-l expression was sodium citrate on ice.